Hyperpolarized (HP) carbon-13 MRI has demonstrated the ability to significantly advance our understanding of disease processes and the great potential to become a cost effective molecular imaging tool for monitoring clinical trial and individual patient management. Our recent first-in-man Phase 1 Clinical Trial utilized unique dissolution DNP instrumentation and methods for [1-13C] pyruvate metabolic imaging. The proposed project is designed to take a major step forward by developing new instrumentation and methods to translate preclinical multi-agent polarization studies into first-in-man dual-agent simultaneous metabolic & perfusion HP MRI studies. This project aims to develop a new MRI approach to characterize cancers based on their genetic/proteomic and perfusion abnormalities and apply this method in preclinical models to obtain the required preliminary data for FDA approval and then to conduct initial human studies. Hyperpolarized (HP) carbon-13 MRI is a powerful new molecular imaging method which uses specialized instrumentation to provide signal enhancements of over 5-orders of magnitude for carbon-13 enriched, safe, endogenous, non-radioactive compounds. Co-polarization of 13C-urea with [1-13C] pyruvate provides not only an internal reference for improved quantitative accuracy, but also a method for simultaneous perfusion measurements without ionizing radiation and without the nephrotoxicity effects of other perfusion contrast agents. This project aims to translate and perform first-in-man HP dual-agent perfusion & metabolic MRI to address a pressing clinical need, specifically improved radiological characterization of prostate cancer aggressiveness and treatment response. New hardware/instrumentation will be designed and constructed to enable dual-agent functionality necessary for first-in-man hyperpolarized perfusion and metabolic imaging human studies. Novel MRI acquisition and analysis methods will also be developed for acquiring rapid high resolution metabolic & perfusion imaging data. The techniques developed in this project will be applied in prostate cancer patients to address a clear unmet clinical need. However, these dual-agent HP 13C MR techniques also have general applicability to advance the clinical research of other cancers and potentially a wide range of pathologies including cardiac, liver, and kidney disease.